摘要
目的:观察复方汉防已对人成纤维细胞增殖和凋亡及合成层粘蛋白的影响,探讨其抗肝纤维化的机制。方法:采用细胞培养法,于细胞培养液中加入不同浓度的药物,再培养48h,行流式细胞仪检测细胞 Fas 基因表达,部分细胞涂片进行 AgNOR 染色,观察 AgNOR 计数变化,培养上清用放免法测定层粘蛋白含量。结果:正常人成纤维细胞 AgNOR 计数为2.12±0.50个核,Fas 基因表达率为15.42%±6.20%,层粘蛋白含量为1.46±0.47mg/L;加入复方汉防已组 AgNOR 计数明显下降,Fas 表达率增加,培养上清层粘蛋白含量减少。结论:复方汉防已能抑制成纤维细胞的增殖和增加凋亡,使合成层粘蛋白减少,这可能是其抗肝纤维化机理之一。
Objective:To explore the anti-fibrotic mechanism of Fufanghanfangji,We observed the proliferation,apoptosis and laminin synthesis in fibroblasts affected by Fufanghanfangji,Method:Different dosage of Fufanghanfangji was added in the cultural fibroblasts.AgNOR,the cell proliferative index was showed with silver dye method.Fas gene protein expression which re- flected cell apoptosis was assessed by flow cytometry.Laminin was assessed by using radioimmunoassay method.Results:In nor- mal cultural fibroblasts,the AgNOR was 2.12±0.50.Fas gene protein positive rate was 15.42±6.20.And laminin content was 1.46±0.47mg/L.The AgNORcount decreased and Fas postive rate increased in fibroblasts cultured with Fufanghanfangji.And the laminin content in supernate decreased in the cultural cell treated with the drug.Conclusions:Fufanghanfangji can inhibit fi- broblasts to proliferate and synthesize extracellular matrix components such as laminin.The drug can also promote cell apoptosis. These may be the anti-fibrotic mechanisms.
